[논문]Improving metabolic efficiency of the reverse beta-oxidation cycle by balancing redox cofactor requirement

Wu, Junjun; Zhang, Xia; Zhou, Peng; Huang, Jiaying; Xia, Xiudong; Li, Wei; Zhou, Ziyu; Chen, Yue; Liu, Yinghao; Dong, Mingsheng

doi:10.1016/j.ymben.2017.11.001

Improving metabolic efficiency of the reverse beta-oxidation cycle by balancing redox cofactor requirement

Metabolic engineering, v.44, 2017년, pp.313 - 324

Wu, Junjun (College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang Road, Nanjing, Jiangsu 210095, China) , Zhang, Xia (College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang Road, Nanjing, Jiangsu 210095, China) , Zhou, Peng (College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang Road, Nanjing, Jiangsu 210095, China) , Huang, Jiaying (College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang Road, Nanjing, Jiangsu 210095, China) , Xia, Xiudong (Institute of Agro-Product Processing, Jiangsu Academy of Agricultural Sciences, Nanjing, Jiangsu 210095, China) , Li, Wei (College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang Road, Nanjing, Jiangsu 210095, China) , Zhou, Ziyu (College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang Road, Nanjing, Jiangsu 210095, China) , Chen, Yue (College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang Road, Nanjing, Jiangsu 210095, China) , Liu, Yinghao (College of Food Science and Techn) , Dong, Mingsheng

Abstract ▼ AI-Helper

Abstract Previous studies have made many exciting achievements on pushing the functional reversal of beta-oxidation cycle (r-BOX) to more widespread adoption for synthesis of a wide variety of fuels and chemicals. However, the redox cofactor requirement for the efficient operation of r-BOX remains unclear. In this work, the metabolic efficiency of r-BOX for medium-chain fatty acid (C6-C10, MCFA) production was optimized by redox cofactor engineering. Stoichiometric analysis of the r-BOX pathway and further experimental examination identified NADH as a crucial determinant of r-BOX process yield. Furthermore, the introduction of formate dehydrogenase from Candida boidinii using fermentative inhibitor byproduct formate as a redox NADH sink improved MCFA titer from initial 1.2g/L to 3.1g/L. Moreover, coupling of increasing the supply of acetyl-CoA with NADH to achieve fermentative redox balance enabled product synthesis at maximum titers. To this end, the acetate re-assimilation pathway was further optimized to increase acetyl-CoA availability associated with the new supply of NADH. It was found that the acetyl-CoA synthetase activity and intracellular ATP levels constrained the activity of acetate re-assimilation pathway, and 4.7g/L of MCFA titer was finally achieved after alleviating these two limiting factors. To the best of our knowledge, this represented the highest titer reported to date. These results demonstrated that the key constraint of r-BOX was redox imbalance and redox engineering could further unleash the lipogenic potential of this cycle. The redox engineering strategies could be applied to acetyl-CoA-derived products or other bio-products requiring multiple redox cofactors for biosynthesis. Highlights The highest metabolic efficiency of r-BOX was achieved. NADH was identified as a crucial determinant of process yield. Fermentative redox balance enabled maximum product synthesis. Two major factors limiting acetate reduction pathway were identified.

주제어

참고문헌 (36)

Science Ajikumar 330 70 2010 10.1126/science.1191652 Isoprenoid pathway optimization for taxol precursor overproduction in Escherichia coli

상세보기
Metab. Eng. Balzer 20 1 2013 10.1016/j.ymben.2013.07.005 Metabolic engineering of Escherichia coli to minimize byproduct formate and improving succinate productivity through increasing NADH availability by heterologous expression of NAD(+)-dependent formate dehydrogenase

상세보기
Metab. Eng. Berrios-Rivera 4 217 2002 10.1006/mben.2002.0227 Metabolic engineering of Escherichia coli: increase of NADH availability by overexpressing an NAD(+)-dependent formate dehydrogenase

상세보기
Nat. Commun. Blazeck 5 3131 2014 10.1038/ncomms4131 Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production

상세보기
Anal. Biochem. Bradford 72 248 1976 10.1016/0003-2697(76)90527-3 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding

상세보기
PLoS Genet. Chou 11 26 2015 10.1371/journal.pgen.1005007 Transhydrogenase promotes the robustness and evolvability of E. coli deficient in NADPH production

상세보기
Metab. Eng. Clomburg 28 202 2015 10.1016/j.ymben.2015.01.007 Integrated engineering of β-oxidation reversal and ω-oxidation pathways for the synthesis of medium chain ω-functionalized carboxylic acids

상세보기
Science Clomburg 355 11 2017 10.1126/science.aag0804 Industrial biomanufacturing: the future of chemical production

상세보기
ACS Synth. Biol. Clomburg 1 541 2012 10.1021/sb3000782 A synthetic biology approach to engineer a functional reversal of the beta-oxidation cycle

상세보기
Nature Dellomonaco 476 2011 10.1038/nature10333 Engineered reversal of the beta-oxidation cycle for the synthesis of fuels and chemicals

상세보기
Nat. Commun. Gajewski 8 14650 2017 10.1038/ncomms14650 Engineering fungal de novo fatty acid synthesis for short chain fatty acid production

상세보기
Appl. Environ. Microbiol. Kim 70 1238 2004 10.1128/AEM.70.2.1238-1241.2004 Effect of overexpression of Actinobacillus succinogenes phosphoenolpyruvate carboxykinase on succinate production in Escherichia coli

상세보기
J. Ind. Microbiol. Biotechnol. Kim 42 465 2015 10.1007/s10295-015-1589-6 Synthesis of medium-chain length (C6-C10) fuels and chemicals via beta-oxidation reversal in Escherichia coli

상세보기
Appl. Microbiol. Biotechnol. Lee 97 2761 2013 10.1007/s00253-013-4750-z Engineering of NADPH regenerators in Escherichia coli for enhanced biotransformation

상세보기
ACS Synth. Biol. Lian 4 332 2015 10.1021/sb500243c Reversal of the beta-oxidation cycle in Saccharomyces cerevisiae for production of fuels and chemicals

상세보기
Metab. Eng. Liu 38 86 2016 10.1016/j.ymben.2016.06.006 C-13 Metabolic Flux Analysis of acetate conversion to lipids by Yarrowia lipolytica

상세보기
Methods Livak 25 402 2001 10.1006/meth.2001.1262 Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method

상세보기
Proteomics Mansour 9 4714 2009 10.1002/pmic.200900161 A proteomic and transcriptomic view of amino acids catabolism in the yeast Yarrowia lipolytica

상세보기
Microb. Cell. Fact. Meng 15 1 2016 10.1186/s12934-016-0536-1 High-yield anaerobic succinate production by strategically regulating multiple metabolic pathways based on stoichiometric maximum in Escherichia coli

상세보기
Nucleic Acids Res. Nakashima 37 e103 2009 10.1093/nar/gkp498 Conditional gene silencing of multiple genes with antisense RNAs and generation of a mutator strain of Escherichia coli

상세보기
Nat. Biotechnol. Qiao 35 173 2017 10.1038/nbt.3763 Lipid production in Yarrowia lipolytica is maximized by engineering cytosolic redox metabolism

상세보기
Appl. Microbiol. Biotechnol. Rutter 99 7359 2015 10.1007/s00253-015-6764-1 Engineering Yarrowia lipolytica for production of medium-chain fatty acids

상세보기
Metab. Eng. Sherkhanov 25 1 2014 10.1016/j.ymben.2014.06.003 Improving the tolerance of Escherichia coli to medium-chain fatty acid production

상세보기
Metab. Eng. Shiba 9 160 2007 10.1016/j.ymben.2006.10.005 Engineering of the pyruvate dehydrogenase bypass in Saccharomyces cerevisiae for high-level production of isoprenoids

상세보기
Microb. Cell. Fact. Tan 16 1 2017 10.1186/s12934-017-0650-8 Improving Escherichia coli membrane integrity and fatty acid production by expression tuning of FadL and OmpF

상세보기
Metab. Eng. van Rossum 36 99 2016 10.1016/j.ymben.2016.03.006 Engineering cytosolic acetyl-coenzyme A supply in Saccharomyces cerevisiae: pathway stoichiometry, free-energy conservation and redox-cofactor balancing

상세보기
Appl. Environ. Microbiol. Vick 81 1406 2015 10.1128/AEM.03521-14 Escherichia coli enoyl-acyl carrier protein reductase (FabI) supports efficient operation of a functional reversal of the beta-oxidation cycle

상세보기
Appl. Environ. Microbiol. Wang 82 7176 2016 10.1128/AEM.02178-16 Engineering of a highly efficient Escherichia coli strain for mevalonate fermentation through chromosomal integration

상세보기
Sci. Rep. Wu 5 14 2015 Enhancing flavonoid production by systematically tuning the central metabolic pathways based on a CRISPR interference system in Escherichia coli
J. Biotechnol. Wu 231 183 2016 10.1016/j.jbiotec.2016.06.007 Stepwise modular pathway engineering of Escherichia coli for efficient one-step production of (2S)-pinocembrin

상세보기
Metab. Eng. Wu 41 115 2017 10.1016/j.ymben.2017.03.012 A systematic optimization of medium chain fatty acid biosynthesis via the reverse beta-oxidation cycle in Escherichia coli

상세보기
Nat. Commun. Xu 4 1409 2013 10.1038/ncomms2425 Modular optimization of multi-gene pathways for fatty acids production in E. coli

상세보기
Proc. Natl. Acad. Sci. USA Xu 113 10848 2016 10.1073/pnas.1607295113 Engineering Yarrowia lipolytica as a platform for synthesis of drop-in transportation fuels and oleochemicals

상세보기
Biotechnol. Bioeng. Zhang 113 2587 2016 10.1002/bit.26021 Optimization of an acetate reduction pathway for producing cellulosic ethanol by engineered yeast

상세보기
ACS Synth. Biol. Zhou 6 1065 2017 10.1021/acssynbio.7b00006 Obtaining a panel of cascade promoter-5 '-UTR complexes in Escherichia coli

상세보기
Nat. Commun. Zhou 7 11709 2016 10.1038/ncomms11709 Production of fatty acid-derived oleochemicals and biofuels by synthetic yeast cell factories

상세보기

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증

Improving metabolic efficiency of the reverse beta-oxidation cycle by balancing redox cofactor requirement

Abstract ▼ AI-Helper

주제어

참고문헌 (36)

이 논문을 인용한 문헌

관련 콘텐츠

AI-Helper ※ AI-Helper는 오픈소스 모델을 사용합니다.

선택된 텍스트